Optimization of the purification of nucleocapsid protein of newcastle disease virus by ultracentrifugation method
Protein purification is a process to remove impurities contains in a complex mixture and may involves a single step or by a combination of several steps. In this study, nucleocapsid (NP) protein of Newcastle disease virus (NDV) was purified using various purification steps from recombinant Escher...
| Main Authors: | , |
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| Format: | Article |
| Language: | English English |
| Published: |
Journal of Pure and Applied Microbiology
2014
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/39237/ http://irep.iium.edu.my/39237/ http://irep.iium.edu.my/39237/1/P_53.pdf http://irep.iium.edu.my/39237/4/Optimization_p821-825.pdf |
| Summary: | Protein purification is a process to remove impurities contains in a complex
mixture and may involves a single step or by a combination of several steps. In this study,
nucleocapsid (NP) protein of Newcastle disease virus (NDV) was purified using various
purification steps from recombinant Escherichia coli. The purification steps involved
ammonium sulphate precipitation, dialysis and sucrose gradient ultracentrifugation. In
this study, the optimized condition for obtaining the highest NP protein yield in the final
step of purification (ultracentrifugation) was determined. Two independent parameters
which are centrifugal force and running time were manipulated and investigated. The
optimum processing condition was determined by employing Face Centred Composite
Design (FCCD) under Response Surface Method (RSM). Based on the almost fully elliptical
plot showed in 2D and 3D response graphs, the optimum condition for obtaining the
highest NP protein yield is near to the centre points of the response surface (condition of
159000xg or 31,000rpm and 5 hours). Although the optimum time to purify the highest
NP protein yield is five hours centrifugation, the NP protein could also be purified after
three hours centrifugation albeit with slight reduction in the yield. Less centrifugation
time is important as pre- and post-centrifugation procedures for sucrose gradient
ultracentrifugation is time consuming and laborious. |
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